Many natural product extracts have been widely used as complementary and alternative medicines (CAM). However, their health benefits and risks have not been subjected to rigorous study, nor have their mechanisms of action been fully understood. The long-term goal of our research program is to validate the medicinal effects of these CAM natural products. The focus of this proposed research is to develop assays that will allow us to identify the molecular targets of CAM natural products of high priority to the NCCAM, and to broadly capture the signatures of biological activities indicative of their complex medicinal effects. Both chemical and biological tools will be used. For the target identification component, we will develop proteomic and cDNA strategies based on both labeled and label-free target identification methods to identify the direct binding targets of resveratrol and Devil's claw, two classes of widely used CAMs. For the signature component, we will use protein profiling to assess the broad biological effects and pharmaco-response pathways of CAM natural products, and develop specific assays and probes to report on the signature response proteins of CAM natural products, thus measuring their in vivo efficacy and facilitating the design of future clinical trials. PUBLIC HEALTH RELEVANCE: Considering the extensive use of natural products as complementary and alternative medicines (CAM) in the US, it is important to study how these natural products work to ensure their proper use. Presently, however, there is little known about the mechanisms of their functions. We propose to identify the molecular targets and drug-response signatures of resveratrol and Devil's claw, two classes of widely used CAMs, to help improve the knowledge of their functions and reduce the risk of their use.